SUMMARY / ABSTRACT Migraine headaches affect up to 15% of people world-wide, including twice as many women as men, and have been called the single most debilitating neurological disorder in the world. Existing first- and second-line treatments including NSAIDS and triptans have, at best, mixed results in the general population as more than half of respondents from a study of >8000 migraine patients reported unsatisfactory pain management with existing treatment options. Clearly, there is a significant clinical unmet need in treating migraine more effectively and a large corresponding (>$4B) market worldwide. Specifically, there is need to develop new classes of drugs with molecular mechanisms of action that are orthogonal to existing therapies that reverse or prevent vasodilation of intracranial vascular smooth muscle cells. Forcyte Biotechnologies is an early-stage bio-pharmaceutical company in Los Angeles that has partnered with advanced high-throughput screening and laboratory automation as well as nanofabrication facilities at UCLA, that is leveraging a microtechnology known as FLECS ? a high- throughput screening (HTS) platform that measures contractility of single-cells in a 384-wellplate format ? to identify and bring to market new compound classes that act on force-generating pathways within cells. This is the first and only reported assay that obtains functional force generation data for single cells, at HTS scales. Our existing programs have focused on treatment resistant asthma and hypertension, and have already had success in discovering novel phenotypic modulators relating to these indications In this proposal, we will build off of our innovative single-cell phenotypic screening platform to develop a functional assay for screening and developing vasoprotective compounds to treat migraine. Our approach replaces older intact artery migraine models, which are extremely informative but very low throughput, with a miniaturized and automated cell-contractility assay implemented in the 384-wellplate format. This will increase throughput from only a few experiments to up to 10,000 experiments per day! Such a transformative improvement will enable us to screen entire compound libraries ? a feat previously impossible ? and greatly improve the probability of successful discovery. Furthermore, the proposed assay would offer high-throughput functional follow-up to other target-based affinity assays that cannot predict phenotypic activity. Once developed, we will focus our assay on high-throughput screens of unique natural product libraries, a rich but relatively underexplored compound space due to throughput limitations of existing methods for evaluating vascular tone and responsiveness. A follow-on Phase II proposal will perform screen an expanded natural product library and perform selectivity counter-screens in related cell types to establish a deep safety profile for the discovered hits, to facilitate transitions to later stages of drug development.